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Ex vivo BIN1/Amphiphysin 2-induced tubulation of membranes, and visualization of Dynamin 2 recruitment to the membrane tubules

External protocol Created on 03 May 2014

Authors

Anne-Sophie Nicot and Jocelyn Laporte

Summary

Membrane remodeling is an important process implicated in several cellular functions including compartmentalization and membrane and protein trafficking. Amphiphysin 2 (BIN1) and dynamin 2 (DNM2) are membrane tubulating proteins, that are mutated in recessive and dominant centronuclear myopathy, respectively (refs 1-4). The overexpression of a muscle isoform of BIN1 (iso 8) in COS-1 cells provokes a striking rearrangement of membranes into long tubules. The large GTPase DNM2 is recruited to these membrane tubules through interaction between its proline-rich domain and the SH3 domain of BIN1. This protocol allows to create and visualise BIN1-induced membrane tubules in cultured cells, and to test the recruitment of DNM2 to the tubules.

Further details

The protocol was published on Protocol Exchange in 2007. To see the entire protocol, click on the source link.

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